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目的探讨MGrl-Ag维持耐药表型与蛋白激酶C的关系, 方法用免疫细胞化学方法检查MGrl-Ag在胃癌SGC7901细胞系及耐药亚系的表达;ELISA及点印迹方法检测胃癌SGC7901细胞系及其耐药亚系细胞培养上清中MGrl-Ag的表达;免疫荧光双标记及共聚焦显微镜技术观察MGrl-Ag与PKCa的相关表达;竞争蛋白结合法检测单克隆抗体MGr-1对PKC活性的影响。结果MGrl-Ag在胃癌SGC7901细胞系及耐药亚系均有表达,以耐药细胞表达明显增多;ELISA及点印迹实验证实在表达MGrl-Ag的细胞培养上清中均可检测到MGrl-Ag;耐药细胞PKCa与MGrl-Ag的相关表达较药敏细胞明显增强;单克隆抗体MGr-1与耐药细胞共同孵育后,耐药细胞PKC的活性减低,以细胞质的PKC降低比较明显。结论 MGrl-Ag可能是一种分泌蛋白,MGrl-Ag维持SGC7901/VCR耐药细胞系的耐药表型可能受PKC信号转导通路的调节。
Objective To investigate the relationship between MGR1-Ag maintain-resistant phenotype and protein kinase C. Methods The expression of MGrl-Ag in gastric cancer SGC7901 cell line and drug-resistant subline was detected by immunocytochemistry. The expression of MGrl-Ag in gastric cancer cell line SGC7901 and drug- The expression of MGrl-Ag in the cell culture supernatant of the drug-resistant subtypes was detected by immunofluorescence double staining and confocal microscopy. The protein expression of MGrl-Ag and PKCa was detected by immunofluorescence double staining and confocal microscopy. Impact. Results MGrl-Ag was expressed in both gastric cancer cell line SGC7901 and drug-resistant subline, and the expression of MGrl-Ag was significantly increased in drug-resistant cells. ELISA and dot-blot experiments confirmed that MGrl-Ag could be detected in the cell culture supernatant of MGrl- The expression of PKCa and MGrl-Ag in drug-resistant cells was significantly higher than that in drug-sensitive cells. After incubated with drug-resistant cells, the activity of PKC decreased and the PKC decreased obviously. Conclusion MGrl-Ag may be a secreted protein. The resistant phenotype of MGrl-Ag in maintaining SGC7901 / VCR resistant cell lines may be regulated by PKC signal transduction pathway.